Diethyl betaaminoethylphosphonate as an algaecide

ABSTRACT

Diethyl betaaminoethylphosphonate can be used to inhibit and/or prevent the growth of undesirable algae, bacteria, fungi, yeast, and other microorganisms. This invention is particularly concerned with the bacteriostatic and bactericidal properties of diethyl betaaminoethylphosphonate compounds against species of Proteus, Salmonella, Shigella, Staphylococcus and Xanthomonas.

United States Patent Kerst et al. June 3, 1975 DIETHYL [56] References Cited BETAAMINOETHYLPHOSPHONATE AS AN UNITED STATES PATENTS ALGAECIDE 3.764.676 10/1973 Kerst et a1. 7l/67 Inventors: Al F. Kerst, Alma, Mich.; John D.

Dour0s, .]r., Gaithersburg, Md.

Assignee: The Gates Rubber Company,

Denver, Colo.

Filed: Mar. 9, 1973 Appl. No.: 339,680

Related [3.8. Application Data Division of Ser. No. 56.940, June 25. 1971, Pat. No.

US. Cl. l. 71/67 Int. Cl A0ln 9/00 Field of Search 7l/67, 86

Primary E.\wniner-James 0. Thomas, Jr. Attorney, Agent, or Firm-Raymond Fink; H. W. Oberg, Jr.; Curtis HQ Castleman, Jr.

[57] ABSTRACT 16 Claims. No Drawings 1 DIETHYL BETAAMINOETI-IYLPI-IOSPHONATE AS AN ALGAECIDE This is a division of copending application Ser. No. 156,940, filed on June 25, 1971. now issued as US. Pat. No. 3,764,677.

BACKGROUND OF THE INVENTION SUMMARY OF THE INVENTION According to the present invention, it has been found that diethyl betaaminoethylphosphonate p CH CH NH C H 0 can be used effectively as an antimicrobial agent. The compound diethyl betaaminoethylphosphonate is well known and may be prepared according to the methods disclosed by A.W. Pudovik and GM. Denisova, Zhur. Obshcewi Khim. 23,263-7(1953) and O Dokiady Akad. Nauk S.S.S.R., 80,658( I951).

DESCRIPTION OF THE PREFERRED EMBODIMENTS The following examples illustrate the antimicrobial qualities of diethyl betaaminoethylphosphonate and describe how these antimicrobial qualities may be utilized in various phases of agriculture, animal husbandry, pharmacology and water treatment technology.

EXAMPLE I Antibacterial and Antiyeast Activity The in vitro effectiveness of diethyl betaaminoethylphosphonate against bacteria and yeast species is established in the following manner. One loopful of each of the investigated bacteria or yeast is transferred from agar slants to 10 ml. of trypticase soy broth and incubated at 37C. for 18 hours. At the end of this period, the bacteria or yeast is seeded into the same medium (1.5-2 percent agar) in which the original inoculum was prepared. The bacteria is then seeded at 1 ml. of inoculum per 250 ml. of medium, which is equivalent to at least 1X 10 cells/m1 as determined by dilutiOn platecount 0r nephelometer readings vs. BaCl Standards. The resultant mixtures are poured into heatresistant sterile petri dishes at a temperature of 45C. Analytical filter paper discs 1.2 cm. in diameter are used for the agar diffusion technique. Each disc is saturated with 0.08 ml. of the solubilized diethyl betaaminoethylphosphonate compound which has been diluted to equal ug./disc and placed on the surface of the hardened agar. The plates are then incubated at 37C. for 18 hours. The activity of the diethyl betaaminoethylphosphonate compounds is established by measuring the zone of inhibition in centimeters. Untreated control plates are used as a basis for comparison and these exhibit a profuse growth of bacteria. The results of these tests are as follows:

Zone of Inhibition Gram positive and Gram in Centimeters negative bacteria Streptococcus Iwmolyric Group A Streptococcus /1!11U1 \!l(' Group B Xanr/zomunasplzuseoll ATCC No. 9563 2.9

Staphylococcus aarem' ATCC No. 2091 Escherichia coli ATCC No, 9637 Erwinla t'arolovora ATCC 9653 Shigt'lla boyzlii ATC C No. 9212 S/iigella lloyllil ATCC No. 9905 3.2 Shlgclla .tonnei MMV 6654 3. S/iigclla flatnerl TYPE 6 MMV 760 S/rigella flawwri TYPE 4 MM\ 6625 Sliigella rlysenlariza' TYPE 2 MMV 6673 Neixseria gonorr/zoeae ATCC No. 19424 Nelsserla intracelluluris Neisseria meningitlrles ATCC No. 13077 Lixlcria manucymgenr'x ATCC No. 15813 0 Proleax valgarilr ATCC No. 9484 Enrinla tarolorara ATCC No. 495

Mvcobacu'riam halyriram ATCC No. 11314 Murvbavleriam jbrlailaln, Debos ATCC No. 4243 Mycobaz'lerium avian:

ATCC No. 19421 Mvcobacrcrimn smegnuais ATCC No. 10143 Mvzohaz'lerimn plllti ATCC No. 11782, phage host Micrococ'cas relragena -Continued Zone of Inhibition Gram positive and Gram in Centimeters negative bacteria Corynebaclerium diplrllreriae ATCC No. 19409 Corynebacterium lraemolylicum ATCC N0. 9345 Diplococcus intracellularix Diplocaccus pneumaniae ATCC No. 6303 Hemoplzilus lremulylicur ATCC No. 10014 Hemoplrilus influenzae ATCC No. 19418 Hemoplzilus paruinfluenzue ATCC No. 7901 Hemoplrilus .ruis

ATCC No. 19417 Hemophilus vaginalis ATCC No. 14018 Brucella aborlus ATCC No. 4315 Brucella melilensis ATCC No. 19396 Brucella suis ATCC No. 4312 Note H hazy zone.

EXAMPLE 11 Secondary Screen Antibacterial Activity Concentration Compound: Diethyl 1000 ug/disc) betaaminoethyl phosplronate MlCROORGANlSM Salmonella sp. ATCC No. 9120 4.1 Salmonella l \'phinzurium miraglia SR] 1 4.4 Salmonella No. 45 MMU 1063 4.4 Salmonella sp. MMU 6674 4.1 Salmonella sp. MMU 6668 4.6

S/u'gella boydu' ATCC No. 9212 5.0 Slzigella boyzlii ATCC No. 9905 5.1 S/rigella flarneri Type 6 MMU 760 5.0 S/rlgella flexm'ri Type 4 MMU 6625 5.1 Slu'gella dyxenteriae Type 2 MMU 6673 5.1 S/ligellu sun/rel MMU 6654 5.0

Pmleus vulgar/s ATCC No. 9484 2.4 H Prmem vulgarix ATCC No. 8427 2.6 H Prawns vulgaris ATCC No. 4699 2.6 H Proteus \ulgaris ATCC No. 6896 2.5 H Proteus r'ulgarix ATCC No. 9920 2.6 H Pmreus rulgurrlr ATCC No. 6897 2.5 H

-Continued Compound; Diethyl Concentration hetuaminoethyl phosphonate 1000 ug/disc) MlCROORGANlSM Proteus r'ulgarlx ATCC No. 12454 2.0 H Pruleux mirahillr ATCC No. 9961 2.3 Proteus morgam'i G951 2.2 Proteus mirabilis G912 2.2 Proteus mirubilis K723 2.2

This data indicates that diethyl betaaminoethylphosphonate compounds can be used to inhibit many important types of diseases. For example, they can be used against:

Bacteria Disease Produced Major cause of bovine mastitis Bacterial endocarditis Cause of scours and secondary cause of bovine mastitis Major cause of fish disease (trout. pike) Numerous animal and plant infections are caused by various pseudomonads. Urinary infections are notable examples.

Various species of Erwr'nia attack commercial crops of carrots, tobacco, potatoes, squash, et cetera.

Various species of Xanlhamonar cause a variety of diseases in plants such as sugar cane. rice, sugar beets, cotton. walnuts, wheat. rye, barley, beans, et cetera.

Sraplrylomccur aureux Slreptomccus faecalir Escherichia coli Proteus r'ulgaris Pxeuzlomonux aeruginoxa Erwinlu carlovora Xanllmmonas p/raseoli The demonstrated antibacterial activity of diethyl betaaminoethylphosphonate compounds against S rap/1- yloc'occus aureus and Escherichia coli is of particular interest to the field of pharamacology since the disclosed activity indicates that these compounds can be formulated as powders, salves, and ointments for administration in the treatment of burns and bacterially induced inflammations such as abscesses, dermatitis, rashes. and the like, particularly in domestic animals.

Although the precise mode of action whereby diethyl betaaminoethylphosphonate inhibits bacteria growth is not completely understood, it is believed that the diethyl betaaminoethylphosphonate compounds of this invention may serve as chemical antagonists; that is, as chemicals which compete with enzymes essential to the development of many bacteria. Since enzymes perform their catalytic function by virtue of their affinity for their natural substrate; any compound resembling a substrate in its chemically critical aspect may also have an affinity for the enzymes. If this affinity is great enough, the analog will displace the normal substrate from the enzyme and will prevent the growth reaction from taking place. It is believed that diethyl betaaminoethylphosphonate has a chemical affinity for an essential site on one enzyme necessary for bacterial growth and life.

The diethyl betaaminoethylphosphonate formulations of this invention can also contain other therapeutically valued supplements such as local anesthetics, irradiated oils, and other medicinal substances. When used for these or similar purposes, this compound may be incorporated in any therapeutically acceptable carrier such as oils, salves and ointments, together with ad juvants comprising surface active agents, detergents, dispersing agents, stabilizers and other modifiers which may facilitate the handling and application of the antibacterial material. In the case of the in vitro applications of the compositions of this invention. it is difficult to predict with precision what in all cases will constitute a therapeutic dose even on a weight basis. Variable factors such as type, duration and severity of infection and mode of administration may be determining factors for the establishment of therapeutic doses.

Those skilled in the art will recognize that the above data indicates that the scope of this invention should not be limited to any particular disease species or to any particular type of animal or plant life. For example, the noted activity of diethyl betaaminoethylphosphonate against Xam/zomonas phaseoli suggests that this compound will also prove to be of value against such other Xanthomonas species as Xanthomonas transluscens, Xanthomonas juglandis Xanthomonas vesicatoria, Xanthomonas barbareae, Xanthomonas pelargonl'i, Xanthomonas alfulfue, Xanthomonas vasculorum, et cetera. Xanthomonas species are known to cause diseases of tomatoes, sugar cane, rice, sugar beets, cottom, walnuts, wheat, rye, barley, and beans. Some of the more noteworthy Xanthomonas related diseases are Xanthomonas vesicatoria (Bacterial Leaf Spot of Tomatoes), Xanthomonas p/zaseoli (Common Bacterial Blight of Bean), Xanthomonas vasculorum (Gumming Disease of Sugar Cane) and Xanthomonas malvacearum (Bacterial Blight of Cotton).

The efficacy of diethyl betaaminoethylphosphonate under simulated field conditions is established by the following tests.

EXAMPLE III Inhibition of Xanthomonas Vesicatoria (Bacterial Leaf Spot of Tomatoes A diethyl betaaminoethylphosphonate composition is prepared for spraying by dissolving it in a suitable solvent such as acetone, methyl alcohol or ethyl alcohol and then diluting the solution to the desired concentration with deionized water containing wetting and dispersing agents. Tomato seedlings, Bonny Best variety, in 7-leaf to 8-leaf growth stage, are mounted on a compound turntable and sprayed at 35 pounds pressure for 50 seconds with diethyl betaaminoethylphosphonate solutions at the concentration indicated below. After drying, treated plants are spray-inoculated at 30 pounds pressure with an aqueous cell suspension of Xanthomonas resicaroria containing five per cent Carborundum and then immediately placed in an incubation chamber maintained at 70F. and 95 percent plus relative humidity. After 40 hours in the incubation chamber, the plants are removed to the greenhouse for further development of the infection. Disease severity is determined by counting the lesions present on 6 to 7 treated leaves. The effectiveness of treatment is determined by direct comparison with inoculated controls. Streptomycin sulfate is used as a reference stan-. dard. The results of these tests are as follows:

at 200 ppm:

EXAMPLE IV Inhibition of Phythopthora Infestans (Late Blight of Tomatoes).

The efficacy of diethyl betaaminoethylphosphonate against Phyt/zopthora Infestans (Late Blight of Tomatoes) is established in the following manner. Bonny Best tomato plants, Lyc'opersicon esculerztum, approximately 5 to 6 weeks old, in five-leaf growth stage, are mounted on a compound turntable and sprayed at 30 pounds pressure with diethyl betaaminoethylphosphonate at the concentrations indicated below. The test compound is prepared for spraying by dissolving it in a suitable solvent (acetone, methyl alcohol, or ethyl alcohol) and diluting to desired concentrations with deionized water containing wetting and dispersing agents. After drying, treated plants are spray-inoculated with a mixed sporangial and zoospore suspension of Phytopthora Infestans and immediately placed in an incubation chamber maintained at F. and percent plus relative humidity. After 40 hours in the incubation chamber, the plants are removed and observed for total infection lesions of the top three leaves. The effectiveness of treatments is determined by direct comparison with inoclated controls. Maneb is used as a reference standard. All units of test include a minimum of three replicates. The results of these tests are as follows.

7: Inhibition at 500 ppm: l8% 7: Inhibition at l000 ppm: 2671 Control (Maneb Inhibition) at ppm: 9771 EXAMPLE V Antifungal Activity The antifungal activity of this compound is established by treating Fusarium oxysporum, Fusarium roseum Rhizopus nigricans, Rhizopus stolonifer, Aspergillus niger and Allernarz'a solani test fungi in the following manner: One loopful of each of the tested viable fungi cultures, spores and mycelia is transferred from an agar slant to an 80 ml. portion of the nutrient broth composed of oatmeal agar, Czapeks, Sabouraud and deionized water to volume. The 80 ml. portion of the fungi and broth is ten placed in a sterile shake flask (300 ml.) and the flask is placed on a rotary shaker for 96 to hours at room temperature. At the end of this incubation time period, 10 ml. of the liquid are homogenized and placed in another sterile shake flask (300 ml.) containing 80 ml. of the above nutrient broth and 60 ppm of diethyl betaaminoethylphosphonate. The flasks are placed on a rotary shaker operating at 240 rpm at room temperature for 3 to 9 days. After this second incubation time, the flasks are taken off and examined for visible fungal growth and mycelia] weights are determined. Untreated controls are used as the basis of comparison and these display profuse fungal growths containing species Fusarium, Aspergillus and Alternaria. The results of these tests indicate that diethyl betaaminoethylphosphonate imparts a substantial degree of inhibition of fungal growth at 60 ppm.

EXAMPLE VI Antifungal and Antiyeast Activity To further define the antifungal activity of this compound the seeded agar plates are prepared by transferring the cultures from slants washed with saline or phosphate buffers to the surface of hardened Sabouraud-Dextrose agar plates. Again as in in the case of Example 1, the diethyl betaaminoethylphosphonate is tested by impregnating filter paper discs (1.27 cm. in diameter) with 0.08 ml. of the solubilized diethyl betaaminoethylphosphonate compounds so as to contain 100 ug/disc and placing them on the surface of the hardened agar. The plates are then incubated at 30C. for 18 hours. The activity of this compound is established by measuring the zone of inhibition in centimeters. Untreated control plates are used as a basis for comparison and these exhibit a profuse fungal growth. The results of these tests are as follows:

Zone of Inhibition Microorganism in Centimeters Aspergillus niger ATCC No. 1004 3.6 Allernaria solam' ATCC No. 6396 3.4 Rhizopus .rlulumfer ATCC No. 10404 3.4 Fusarium Qryqxlmm UFCC 1122 2.2 Candida albicunx EXAMPLE Vll Secondary Fungal Screen A secondary screen using the techniques of Example Vl produces the following results at the diethyl betaaminoethylphosphonate concentration indicated:

Concentration 100 ug/disc) -Continued Concentration ug/disc) MlCROORGANlSM P/mma piumunloruru ATCC No. 12569 4.0

Pzu'z'ilunrvr'vs variuli ATCC No. l 1 14 4.1

Nigmt mra spline/it'd Penicillium rubrum ATCC No. 10520 4.2 Penicillin/n lmlatum OU 4.3

Beuuvaria busxianu MV 1341 4.1 Bcuuvaria renal/a MV 1919 4.2

Torn/u lu'rgeri OU 4.2

Mmwrporum apim'pvrmum OU 4.1 Allernnricl .roluni ATCC No. 639 3.4 H

Georriclmm sp. OU 2.1 H

Verlicillium ulbo-ulrmn ATCC No. 10833 3.9

'Ii'ichuphywn I)l('Illflgl'0p/1 \Ie.\ ATCC No. 9129 4.3 Triclzophylon 1uenlugrupliyrcs ATCC No. 8215 4.1 'll'icliopliylun mnxuruns ATCC No. 10217 4.2

Cermx mru helicola ATCC No. 12825 4.0

Pyl/zium urrhenomunvr ATCC No. 12531 4.1

Ht Imirzllmxpnrium oryzav ATCC No. 11000 4.2

Comments:

H hazy tr trace It will also be recognized by those skilled in the art that other protectant, systemtic and eradicant procedures may provide detection of other biological activities. Pathogens representative of Phycomycetes, Ascomycetes, Basidiomycetes and the Fungi Imperfecti may provide indices of other fungicidal activity. Additional pathogens and appropriate host organisms may well afford other opportunities to further define the degree and spectrum of the activity disclosed in this invention. Since no firm rules of procedure can be laid down for the sequence of such evaluations or for the choice of pathogens, diethyl betaaminoethylphosphonate must be considered on the basis of its demonstrated performance in such primary evaluations and then progressively judged in subsequent studies. A wide range of pathogens. representative of economically important diseases, can be used to help define this compound's biological activity and to assure high degrees of success under field conditions. The following disease organisms, crops and reference standards may be used in such evaluations:

Disease -Continued Disease Organism Reference Compound Downy Mildew of Sugar Beet Downy Mildew of Lima Bean Bean Rust Powdery Mildew of Wheat Powdery Mildew of Apple Powdery Mildew of Roses Powdery Mildew of Cantalope Leaf Spot of Wheat Early Blight of Tomato Rice Blast Disease Cercospora Leaf Spot of Sugar Beets Septoria Leaf Spot of Celery Apple Scab Common Bacterial Blight of Bean Uromyct'i' pltaseoli var. Iypiw Erysiphe gruminis Pollospliaeru lt'ttc'olricliu Spliat'rollier'a pmiiiosu var. rosin Erysip/u' cic'lmrucearum Helmi'nrlmsporium Salii'imi Altvrnariu solum' Piri'r-ularizi uryzae Cercuspora bericolu Seplorin api'i-grai'culenri's Venluria inaequali's X anr/izimomis plmseo/i Karathane Karathane Maneb Karathane Karathane Karathane Karathane Maneb Maneb Blasticidin Maneb Maneb Cyprex Streptomycin Sulfate nized that the above formulations with slight modifications may be used in the field of animal husbandry as dusting powders and salves.

Where it is desired to use the aforementioned wettable powders or liquid formulations, either emulsified, dispersed or suspended in water or other fluids, one or more of the class of materials herein referred to as adjuvants can also be incorporated into the powder, dust or liquid formulation. These adjuvants comprise surface active agents, detergents, wettable agents, stabiliz- BrownR t fSr Fruits 0 ers, dispersing agents, suspending agents, emulsifying GrgyvMold to]! [fruit 3 Munch agents, Spreaders, stickers and conditioning agents gengl gfs g i D M b erally. To their modifying characteristics these adju- I: Vegetable Rm vants may facilitate handling and application and infreg g ht g immigrant rligilulimi Maneb quently enhance or potentiate the diethyl betaaminoet'nici iimi imlicimi Karathane Bacterial pis f Rmdommm WWW Capmn thylphosphonate compositions of this invention in their 5 F pbiological activities by mechanisms which are freactem] Soft 91pm" quently not well understood. A satisfactory but not ex- In their plant protection aspects, the diethyl betaaminoethylphosphonate compounds of this invention may be used in the manner known to the organophosphorus crop protection art; that is, they can be made up in solid or liquid formulations. Examples of solid formulations are dust, wettable powders, granules and pellets. As a dust, diethyl betaaminoethylphOS- phonate compounds may be dispersed in powdered solid carriers such as talc, soaps, soapstone, attalpulgus clayas well as other finely divided solids known to the dusting art, When formulated as wettable powders, the active component may be employed in conjunction with inert fillers which may be of the clay type carrier or non-clay type, in conjunction with various combinations of wetting agents and emulsifiers which permit the adaptation of the concentration as a free-flowing pO der for dispersion in the field.

Each of these carriers may in turn contain other carriers or extenders which are ordinarily non-reacting or inert substances such as sand, clays, talc, sawdust, alkaline earth carbonates, oxides, phosphates and the like as well as diatomaceous earth, micas or other suitable materials. When liquid formulations are desired, liquid extenders, dilutants or carriers of a non-reactive nature may be utilized. Examples of such materials are alcohols, ketones, glycols, aromatic hydrocarbons, petroleum fractions such as Octane and various other distillates. From these considerations, it will also be recoghaustive list of these adjuvants appears in Soap Chemical Specialties, volume 31, no. 7, page 61; no. 8, pages 38-61; no. 9, pages 52-67; and no. 10, pages 38-67 (1955). See also, bulletin no. 607 of the Bureau of Entomology and Plant Quarantine of the US. Department of Agriculture.

An additional advantage of diethyl betaaminoethylphosphonate is its compatibility with a variety of other bactericidal and fungicidal materials. For example. it may be convenient to combine this compound with one or more other adjuvants. carriers. pesticides. biocides or fungicides of various structures. For example, diethyl betaaminoethlphosphonate fungicidal inhibitors may be combined with insecticidal materials such as chlordane, benzene hexachlorides, DDT. DDD, the insecticidal carbamates, polychlorinated terpenes. parathions, methoxychlor, insecticidal phosphates, phosphorothioates, phosphorodithioates and with fungicides such as sulphur, quinones, dodecylgaunidine and metal dimethyldithiocarbamates.

There are many other considerations which may make some methods of application more favored than others. These considerations may include the type of organisms on which the compound is to be administered, the degree of activity, the degree of activity to ward the particular organism, and side effects. Also to be considered is the cost of production and the characteristic solubility of diethyl betaaminoethylphosphonate in the cairier material.

The broad spectrum of antifungal activity afforded by this compound can also be utilized in the formulation of disinfectant solutions, paints, coatings, films and polymeric materials in order to protect against disease and rot caused by various fungi species. When used as a disinfectant, suitable formulations may be prepared by mixing the compound with an emulsifying agent in the presence of organic solvents and then diluting it with water to form an aqueous emulsion containing the diethyl betaaminoethylphosphonate. Suitable emulsifying agents include, e.g. alkylbenzenesulfonates, polyalkene glycols, et cetera. Aqueous emulsions of diethyl betaaminoethylphosphonate are particularly suited for use in disinfectant solutions used in washing hospital floors and walls. The following examples further illustrate the antifungal properties of diethyl betaaminoethylphosphonate.

EXAMPLE VIII Preparation of a Vinyl Coating Resistant to Mildew Deterioration EXAMPLE IX Preparation of Plasticizers Resistant to Mildew A commercial thermoplastic monomer is divided into portions which are treated as follows.

Portion I:

To the first portion is added 2 percent by weight of diethyl betaaminoethylphosphonate and percent by weight of dimethylnaphthalate as plasticizer. The monomer is polymerized and molded into 3-inch diameter discs, one-fourth inch in thickness prior to testing.

Portion 2 To this portion is added 2 percent by weight of diethyl betaaminoethylphosphonate and 10 percent by weight of butyl isodecylphthalate as plasticizer. The monomer is polymerized and molded as above.

Portion 3 This portion is the untreated control of Portion 1 containing no fungal inhibitor but 10 percent by weight of dimethylnaphthalate as plasticizer. Again, the polymerization and molding are identical.

Portion 4 This portion is the untreated control of Portion 2 containing no fungal inhibitor but 10 percent by weight of butyl isodecylphthalate as plasticizer. The polymer ization and molding are described above.

The two plasticizers were chosed on the basis of their known susceptibility to Fusarium attack under high humidity and temperature conditions.

EXAMPLE X Test of Vinyl Coatings and Plasticizers For Fungal Resistance The vinyl coated articles and controls of Examples VIII, IX are placed in an air-tight high temperature and high humidity chamber maintained at F. and percent humidity to stimulate tropical temperature and humidity conditions. After a months exposure the vinyl coated articles treated with diethyl betaaminoethylphosphonate inhibitor are only slightly attacked by rot while the articles not treated are well rotted. The two untreated control polymer discs are examined and found to be blackened and mildew rotted. Isolates of Aspergillus, Fusarium and known species of yeasts are prepared from the deteriorated discs. The discs containing the diethyl betaaminoethylphosphonate fungal inhibitor are not adversely affected.

EXAMPLE XI Algaecidal Properties of Diethyl Betaaminoethylphosphonate The effectiveness of this compound against various algae species is established in the following manner. Algal cultures representing Scenedesmus. Chlorella, Plectonema, Anacystis, Ankistrodesmus, Anabaena, Synura, Oscillatoria, Coccochloris, Chlamydomonas and Lyngbya are each maintained in Chu no. 10 Broth Medium (Calcium nitrate, 0.040 grams; Potassium phosphate, 0.010 grams; Magnesium sulphate, 0.025 grams; Sodium carbonate, 0.020 grams; Sodium silicate, 0.025 grams; Ferric citrate, 0.003 grams; Citric acid, 0.003 grams; and deionized water, 1,000 ml.) in the presence of sunlight. Hardened Chu no. 10 agar plates are inoculated with swabs saturated with the respective algae broth cultures. The diethyl betaaminoethylphosphonate is tested by impregnating filter paper discs (1.27 cm. in diameter, no. 740-E, Schleicher and Schuell, Keene, New Hampshire) with a solution of the diethyl betaaminoethylphosphonate at the concentrations indicated. The saturated filter discs are placed on the surface of the seeded agar plates and the optimum growth temperature of 20 to 27C. is maintained. Untreated control plates are used as a basis for comparison and these exhibited a profuse growth of algae. The results of these tests are expressed as inhibition zone diameters.

ATC C 11469 4.0 2.1 tr Lc nn'im'lili' ()sillumriu Gmnp/iunemu Plt't'lom'mu HOIUIIHH .\'i1:.\'('hi l Lynghya C/Iltl/H)(lH1UI|tlS Taft EEC 172 l8 If U Cl!IllI)l)'ll()/7t)ll' Antlers-11's iiidulanx H) Taft EEC l34 0 O O SL'RFACE WATER ALGAE GENERA AH/(iSIIOdL'SIHHS var. Aclinuxlrimi EIHISII'IUH Zygm'mu at'icu/urix .N'uduluriu (imiiimi Sltllll'UIlt'fa' Taft EEC 28 0 U Cuz'lusll'um Dexmizlium Spliuvlm'vi'lis' Anubaena z'ulenula Eng/mu! Pediustrum SPHLYIKYHHLY SRl EEC 315 4.2 3.0 l.l Mivructim'um Emlm'inu ()oqvxtix S \-nuru ulre/Iu l Mongmliu (in!iip/mxp/im'riu Ul (Ll 5.4 4.0 ()rt'illamri'u mnit'ii RESERVOIR ALGAE GENERA OU 5,0 4.] 2 l Clulru Amlnuim'lla ('om ii'u wgmi (occur/Hurts eIe/uuis Plmrmidi'um Telruspor'rl Bill/HT'IIOSIHI'HHHII SRl 4. 3 7 .4 l'lorliri'x Aclilizuillwx Cymhellu (Ii/um \(1l)IIll)HUS rmliuii ('Imlop/mru .S'ligeoclunium Bulhoc'lmt'u' UA 4.0 2.0 tr Gum zlimienm Lynglrvu Dra umiuldiu Lynghyu sp. Taft EEC 166 3.9 3.7 2 2 mum, The scope of this invention should encompass the use h me of this compound in waters of all types such as lakes, 5 rivers, ponds, streams, reservoirs. swimming pools and oceans as well as recirculatin industrial waters. These As was the case with the bacteria. it should be recogb g compounds can e used to prevent the initial occur nized that the scope of this invention should not be limd I F I l rence of algae or they can be used on bodies of water ite to any particu ar species. or instance, the algae I I y b with algae already looming. The iethyl betaaminoe cidal activity of this compound against (lilorella vulgue thylphosphonate compounds of the present invention ris. Chlorella ellzpsozdea and Chlorella arrwwulosa sugare also advantaceous in that the are de radable with gests that the compound will also prove to be of value f h d c d d y b g h none 0 t e egra ation pro ucts eing '[OMC to is against other species such as Clilorella i-ariegaru, et cetand most fish food Organisms at algae killing concentra era. Similar possibilities exist for species of the other tions nera whose activit was shown to b arr Ound It shouldybe reco nized aleso ii i Another important advantage of diethyl betaaminoec0 g er pthylphosphonate compounds in their algaecidal applipropriate algae genera may well afford additional opf h d h d I cations is that they can be made up in solid or liquid portumFles urt t e egiee d.nd.spectr.um formulations. Examples of solid formulations are dust, of the algaecidal activity disclosed in this invention. wettable powders granules and pellets. Solid formulw Smce fig z z i gjgfis gig gfgt igiz f the 40 tions, particularly floating solid formulations, may be quenc g k v c Ion among preferred in combating algae which grow on surface the molie than 20O00 algae i l' waters. As dusts, these compounds may be dispersed in betaaminoethylphosphonate compounds of this invenpowdered Solid Carriers such as talc Soap goapstone th th g g fs s i l I attapulgus clay, as well as other finely divided solids. S Ta 8 per e p a f When formulated as wettable powders, the active dithen progressively udged in subsequent studies. These ethyl betaaminoethylphosphonate component may be evaluations should include but not be limited to the folemployed in conjunction with inert fillers which may be lowmg algae genera of the clay type carrier or non-clay type in conjunction with various combinations of wetting agents and emulsifiers which permit adaptation as a free flowing pow- TASTE AND ODOR C AUSING ALGAE GENERA der. Each of these carriers may in turn be combined TU with other carriers which are ordinarily non-reacting or M inert substances such as sand. clays. talc. sawdust. alka- Z e zy P- if gw' fw'dvri/w line earth carbonates. oxides. phosphates and the like.

\'l I'Ol I('l \(l!l (I'll IIUH Hlll'tl t I g (hcloxp/uuvnun as well its diatomaceous earth. micas or other suitable materia s.

CLEAN WATER ALGAE GENERA y x (Inga/Him Mcrlzvnwlwdm Mmdm When liquid formulations are dCSlfLd. liquid extend Pilillll/(ll'izl A /mmn/aw (/lrumulimr ers or carriers of a non-reactive nature may be utilized. Z "0' f 1111mm These compositions should contain approximately 0.1 Rlmllunmnax .\tl\'!(ll[tl Mum-minim 20 b A Surirvllu (lulmuui'ip/mn Lt'llkllllil to Percent y gh and preferably O.l t0 3 percent r/"1141a Mirna-arias ar aand most preferably 0.5 to 2 percent of the active in- CIII' \.\NI.'(I(L'H.\ (ululliru .Wmvrulvmv gredient Solvents hich m ly be uged in the prepara AII/(iSlIr/[IUJHIILV c l g r v v tion of such compositions would include alcohols. kemihmm Fll-TER (1M1 ZS Z tones. glycols. mineral spirits and aromatic solvents C/II'IKM'UL'L'IIA' 'l'uln'lluriu 'l'mclit'liunmius ,5 such as henzeney nitrobenzenedimethylfor' g fy gn "fi gy' mide. Furthermore, to assist in the rapid and complete \III F U ('(M'II'U ll Hit (I I ("lilurvllu ('yr'lnlt'llu Din/mm! dlspersmnm wiltefyslems the? compofiltlons Synvdm A'rn-it-u/u lmgilm'ui also contain approximately 5 to 30 percent by weight and preferably to percent by weight of surfaceactive agents. Suitable surface-active agents include sodium dialkyl sulphates, sodium alkylbenzenc sulfonates. sodium carboxylates and the non-ionic surfactants such as ethoxylated fatty acid alcohols and amines.

Diethyl betaaminoethylphosphonate is compatible with a wide variety of other algaecidal, bactericidal and fungicidal materials. For example, it may be convenient to combine this compound with one or more of the other bactericidcs, fungicides or algaecides. For example, common fungicides or bactericides such as sulphur, inorganic salts such as copper sulphate. activated colloidal silver compounds. copper naphthenate and zinc acetate, as well as substituted hydrocarbons and ammonium compounds, may be employed.

Other considerations may make some methods of application and use of this compound more favored than others. These considerations may include the type of organisms on which the compound is to be administered, the degree of activity, the degree of inhibition toward the algae organism and possible side effects. Also to be considered is the cost ofproduction and the characteristic solubility of diethyl betaaminoethylphosphonate in the carrier compounds.

In their algaecidal aspects the applicants have discovered that diethyl betaaminoethylphosphonate compounds are active algaecides at relatively low concentrations. For example, it has been discovered that these Compounds have algaecidal activity at concentrations as low as 0.1 ppm. The amount of diethyl betaaminoethylphosphonate added to the water will, of course, vary depending upon such factors as the type of algae present, the nature of the body of water, i.e. flowing stream versus small lake, et cetera, and the inherent ability of the body of water to support algae growth. This inherent ability in turn depends upon such factors as exposure to sunlight. pH, nutrient capabilities, and the like. In most cases, however, the concentration of this compound required to kill or inhibit growth of algaes will vary from 0.1 to 10 ppm, with the preferred amount being in the range of 0.8 to 5 ppm.

This compound can be added to the water according to conventional techniques for algaecide applications. When treating a lake or body of water which is relatively calm, the conventional procedure is to spray an aqueous solution of the algaecide over the surface of the water. The active ingredient generally will be predissolved in the types of solvents previously mentioned. In the case of moving water, such as in water treatment plants or industrial facilities, the algaecide can be added to the water in small amounts at periodic intervals. For economic reasons, volume usages such as in lakes, streams, reservoirs, as distinguished from specialized uses such as in aquatic gardens. and industrial applications, the concentrations of the diethyl betaaminoethylphosphonate algaecides probably will not be more than 0.8 to 5 ppm of the water containing the algae.

Having thus disclosed our invention, we claim:

1. A method of killing and inhibiting the growth of algae which comprises contacting said algae with diethyl betaaminoethylphosphonate in an amount effective to kill and inhibit the growth of said algae.

2. The method of claim 1 wherein the algae are contacted with growth inhibiting amounts of diethyl betaaminoethylphosphonate together with acceptable carriers in an amount effective to kill and inhibit the growth of said algae on an plant infected with said algae.

3. The method of claim 1 wherein the algae is Scenedesmus.

4. The method of claim 1 wherein the algae is Chlorella.

5. The method of claim 1 wherein the algae is Plectonema.

6. The method of claim 1 wherein the algae is Anabaena.

7. The method of claim 1 wherein the algae is Syrura.

8. The method of claim 1 wherein the algae is Oscillatoria.

9. The method of claim 1 wherein the algae is Coccochloris.

10. The method of claim 3 wherein the Scenedesmus is selected from the group consisting of Scenedesmus busilensis and Scenedesmus obliquus.

11. The method of claim 4 wherein the Chlorella is Clilorellu vulgarl's, C/zlorella ellipsoidal and Clzlorella pyrenoidosa.

12. The method of claim 5 wherein the Plectonema is Plectonema notation.

13. The method of claim 6 wherein the Anabaena is Anabaena carenula.

14. The method of claim 7 wherein the Synura is Synura ulvella.

15. The method of claim 8 wherein the Oscillatoria is Oscillatoria curneti.

16. The method of claim 9 wherein the Coccochloris is Coccochloris ("lCbllllS. 

1. A METHOD OF KILLING AND INHIBITING THE GROWTH OF ALGAE WHICH COMPRISES CONTACTING SAID ALGAE WITH DIETHYL BETAAMINOETHYLPHOSPHONATE IN AN AMOUNT EFFECTIVE TO KILL AND INHIBIT THE GROWTH OF SAID ALGAE.
 1. A method of killing and inhibiting the growth of algae which comprises contacting said algae with diethyl betaaminoethylphosphonate in an amount effective to kill and inhibit the growth of said algae.
 2. The method of claim 1 wherein the algae are contacted with growth inhibiting amounts of diethyl betaaminoethylphosphonate together with acceptable carriers in an amount effective to kill and inhibit the growth of saId algae on an plant infected with said algae.
 3. The method of claim 1 wherein the algae is Scenedesmus.
 4. The method of claim 1 wherein the algae is Chlorella.
 5. The method of claim 1 wherein the algae is Plectonema.
 6. The method of claim 1 wherein the algae is Anabaena.
 7. The method of claim 1 wherein the algae is Syrura.
 8. The method of claim 1 wherein the algae is Oscillatoria.
 9. The method of claim 1 wherein the algae is Coccochloris.
 10. The method of claim 3 wherein the Scenedesmus is selected from the group consisting of Scenedesmus basilensis and Scenedesmus obliquus.
 11. The method of claim 4 wherein the Chlorella is Chlorella vulgaris, Chlorella ellipsoidea and Chlorella pyrenoidosa.
 12. The method of claim 5 wherein the Plectonema is Plectonema notatum.
 13. The method of claim 6 wherein the Anabaena is Anabaena catenula.
 14. The method of claim 7 wherein the Synura is Synura ulvella.
 15. The method of claim 8 wherein the Oscillatoria is Oscillatoria corneti. 